Advancing biological and clinical applications of genomic Minimal Residual Disease detection in AML

推进基因组微小残留病检测在 AML 中的生物学和临床应用

基本信息

批准号：
9763499
负责人：
Sami Nimer Malek
金额：
$ 41.69万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9763499
关键词：
Acute Myelocytic Leukemia Adult Affect Age Alleles Allogeneic Bone Marrow Transplantation Biological Biological Assay Biological Markers Blast Cell Bone Marrow Bone Marrow Cells Cell Separation Cells Cessation of life Chemotherapy-Oncologic Procedure Clinical Clinical Management Complement Consolidation Therapy Cytogenetics Data Decision Making Detection of Minimal Residual Disease Development Diagnosis Disease remission Foundations Future Gene Frequency Gene Mutation Genes Genomics Hematopoiesis In complete remission Individual Maintenance Maintenance Therapy Marrow Measurement Measures Modeling Monitor Mus Mutate Mutation Mutation Analysis Outcome Patients Phenotype Preleukemia Prognostic Factor Publishing Recurrent disease Relapse Residual Neoplasm Residual Tumors Residual state Risk Sampling Source Techniques Therapeutic Therapeutic Agents Time Tumor Debulking Validation Xenograft procedure age related base chemotherapy clinical application cohort digital disorder risk genetic variant improved insight interest leukemia leukemic stem cell mutant novel novel therapeutic intervention novel therapeutics parkin gene/protein predict clinical outcome prognostic prognostic assays prognostic value single-cell RNA sequencing survival outcome transcriptome transcriptome sequencing

项目摘要

ABSTRACT Acute myelogenous leukemia (AML) affects more than 20,000 adult patients in the US per year causing greater than 11,000 untimely deaths. Standard AML therapy comprises chemotherapy induction to achieve leukemia debulking, followed by a few cycles of consolidation chemotherapy alone and/or followed by allogeneic bone marrow transplantation. This general therapeutic approach to AML is risk-adapted relying principally on well-established prognostic factors, including the type of AML, age, gene mutations and cytogenetic results. Despite best efforts, AML remains incurable in the majority of afflicted patients. A principle barrier to AML cure is disease relapse despite achieving a clinical complete remission (CR) following standard chemotherapy regimens. Of substantial interest therefore, is the identification and accurate measurement of residual AML that persists during remission in AML patients, as such residual disease is likely the source of relapse. Several important questions about AML relapse are only in early stages of satisfactory answers, including i) how to best measure minimal residual disease (MRD) in AML, ii) whether all MRD is indeed caused by residual disease or rather as suggested by recent data comprises a mixture of leukemia, pre- leukemia and age-related clonal hematopoiesis, iii) what constitutes the cellular source of relapse in AML, iv) if such relapse-causing cells can be better targeted using novel therapeutic approaches, and v) under what circumstances is MRD prognostic and useful for MRD risk-adapted AML clinical management. Given lack of well-suited techniques to reliably answer the relevant questions detailed above, we have optimized droplet digital PCR (ddPCR), a novel ultra-high sensitivity assay for detecting genomic MRD in AML. In a detailed recently published study (Parkin et al, JCI 2017) we found that AML frequently relapses from rare cells residing in remission marrows that based on mutation analysis resemble AML blast cells detected at diagnosis. We demonstrated the feasibility for detecting AML-associated gene mutations at allele frequencies as low as 0.002% and have also provided important novel prognostic insights. In this application, we are proposing complete characterization of mutational MRD and aberrant cellular clusters in AML remission bone marrows using a combination of sophisticated cell sorting and single cell transcriptome analyses complemented with mouse xenografting and ex vivo colony forming assays. Anticipated findings will improve the functional characterization of cells that carry AML-associated gene mutations in an attempt to identify and better characterize the source(s) of AML relapse. Using AML samples from two clinical validation cohorts, we will define the prognostic utility of ddPCR-based MRD assessments. Overall, data will lay the foundation for future real-time genomic MRD-guided clinical AML trials aiming at monitoring and improving consolidation and maintenance therapy and ultimately survival outcome in AML.

抽象的急性髓性白血病 (AML) 每年影响美国 20,000 多名成年患者，造成更大的损失超过 11,000 人过早死亡。标准 AML 治疗包括化疗诱导以实现白血病减灭术，然后进行几个周期的单独巩固化疗和/或随后进行同种异体骨髓移植。这种针对 AML 的一般治疗方法是根据风险进行调整的主要基于既定的预后因素，包括 AML 类型、年龄、基因突变和细胞遗传学结果。尽管尽了最大努力，大多数患者的 AML 仍然无法治愈。一个尽管在以下情况下实现了临床完全缓解 (CR)，但 AML 治愈的主要障碍是疾病复发标准化疗方案。因此，重要的是识别和准确测量 AML 患者缓解期间持续存在的残留 AML，因为此类残留疾病很可能复发的根源。关于 AML 复发的几个重要问题仅处于令人满意的早期阶段答案，包括 i) 如何最好地测量 AML 中的微小残留病 (MRD)，ii) 是否所有 MRD 均适用确实是由残留疾病引起的，或者更确切地说，正如最近的数据所表明的那样，包括白血病、预白血病和年龄相关的克隆造血，iii) AML 复发的细胞来源是什么，iv) 如果使用新的治疗方法可以更好地靶向此类引起复发的细胞，以及 v) 在什么情况下情况是 MRD 的预后因素，对于 MRD 风险适应的 AML 临床管理很有用。鉴于缺乏合适的技术来可靠地回答上述相关问题，我们有优化的液滴数字 PCR (ddPCR)，一种新型超高灵敏度检测方法，用于检测基因组 MRD 反洗钱。在最近发表的一项详细研究（Parkin 等人，JCI 2017）中，我们发现 AML 经常复发来自缓解期骨髓中的稀有细胞，基于突变分析，这些细胞类似于 AML 母细胞诊断时发现。我们证明了检测 AML 相关等位基因突变的可行性频率低至 0.002%，并且还提供了重要的新颖的预后见解。在此应用中，我们提出突变 MRD 和异常细胞的完整表征使用复杂的细胞分选和单细胞相结合，在 AML 缓解骨髓中进行聚类转录组分析与小鼠异种移植和离体集落形成分析相补充。预期的研究结果将改善携带 AML 相关基因突变的细胞的功能特征尝试识别并更好地描述 AML 复发的根源。使用来自两个临床的 AML 样本在验证队列中，我们将定义基于 ddPCR 的 MRD 评估的预后效用。总体而言，数据将为未来实时基因组 MRD 指导的临床 AML 试验奠定基础，旨在监测和改善 AML 的巩固和维持治疗以及最终的生存结果。